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  Solution structure and functional investigation of human guanylate kinase reveals allosteric networking and a crucial role for the enzyme in cancer.

Khan, N., Shah, P. P., Ban, D., Trigo-Mouriño, P., Carneiro, M. G., DeLeeuw, L., et al. (2019). Solution structure and functional investigation of human guanylate kinase reveals allosteric networking and a crucial role for the enzyme in cancer. The Journal of Biological Chemistry, 294(31), 11920-11933. doi:10.1074/jbc.RA119.009251.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0004-AFFC-C Version Permalink: http://hdl.handle.net/21.11116/0000-0004-AFFE-A
Genre: Journal Article

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 Creators:
Khan, N., Author
Shah, P. P., Author
Ban, D., Author
Trigo-Mouriño, P., Author
Carneiro, M. G., Author
DeLeeuw, L., Author
Dean, W. L., Author
Trent, J. O., Author
Beverly, L. J., Author
Konrad, M.1, Author              
Lee, D., Author
Sabo, T. M., Author
Affiliations:
1Research Group of Enzyme Biochemistry, MPI for biophysical chemistry, Max Planck Society, ou_578612              

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Free keywords: enzyme kinetics; enzyme mutation; guanylate kinase; hGMPK; lung cancer; non-synonymous single nucleotide variants (nsSNV); nuclear magnetic resonance (NMR); protein structure; solution structure
 Abstract: Human guanylate kinase (hGMPK) is the only known enzyme responsible for cellular GDP production, making it essential for cellular viability and proliferation. Moreover, hGMPK has been assigned a critical role in metabolic activation of antiviral and antineoplastic nucleoside-analog prodrugs. Given that hGMPK is indispensable for producing the nucleotide building blocks of DNA, RNA, and cGMP and that cancer cells possess elevated GTP levels, it is surprising that a detailed structural and functional characterization of hGMPK is lacking. Here, we present the first high-resolution structure of hGMPK in the apo form, determined with NMR spectroscopy. The structure revealed that hGMPK consists of three distinct regions designated as the LID, GMP-binding (GMP-BD), and CORE domains and is in an open configuration that is nucleotide binding-competent. We also demonstrate that nonsynonymous single-nucleotide variants (nsSNVs) of the hGMPK CORE domain distant from the nucleotide-binding site of this domain modulate enzymatic activity without significantly affecting hGMPK's structure. Finally, we show that knocking down the hGMPK gene in lung adenocarcinoma cell lines decreases cellular viability, proliferation, and clonogenic potential while not altering the proliferation of immortalized, noncancerous human peripheral airway cells. Taken together, our results provide an important step toward establishing hGMPK as a potential biomolecular target, from both an orthosteric (ligand-binding sites) and allosteric (location of CORE domain-located nsSNVs) standpoint.

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Language(s): eng - English
 Dates: 2019-06-142019-08-02
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1074/jbc.RA119.009251
 Degree: -

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Title: The Journal of Biological Chemistry
Source Genre: Journal
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Pages: - Volume / Issue: 294 (31) Sequence Number: - Start / End Page: 11920 - 11933 Identifier: -